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  ANTIBODIES  

(anti-Human and others as indicated)

RDI Divison of Fitzgerald Industries Intl  offers a wide line of  antibodies. Since no one antibody works best for all applications (neutralization, blotting, ELISA, etc), we offer many different types of antibodies to help solve this problem. Please inquire for other applications or types of antibodies not listed below.


-See other epitope tagging antibodies including maltose binding protein, HA-Tag,  Myc-Tag,   VSV-G-Tag, Thyoredoxin, SjGst  ,Green Fluorescent Protein , GLU-GLU,   KT3,  cmyc, au1, au5, T7, V5
Thioredoxin (see below 4 antibodies)

Mouse (Monoclonal) anti Thioredoxin (trx)

cat# RDI-THIORDabm   $500.00/100ug

Packaging: lyophilized antibody. Reconstitute with distilled water to yield 1.0mg/ml IgG in PBS pH 7.2 with 1% sucrose and 0.1% sodium azide.

Isotype: mouse monoclonal IgG1k

Clone: 2C9

Immunogen: E. Coli thioredoxin (trxa) 109aa

Specificity: This antibody detects the E. Coli trx and trx-tag- fusion protein on immunoblotting. It detects as 12kDa band of E Coli trxa by immunoblotting on total cell lysate from induced E. Coli harboring Trx-expression vector such as pTrxFus(Invitrogen) without any insert. -western blot (approx 1ug/ml with chemiluminescence detection)


ref: -La Vallie, E.R. et al, Bio/Technology, 11:187(1993)

       -Novy,R et al, inNovations, 3:7(1995)

Storage: Lyophilized antibody is stable for at least 1 year from date of shipment when stored at 4 DEG C. Reconstituted antibody should be stored at 4 DEG C short term or aliquoted and frozen at -20 DEG C. Avoid frequent freeze thaw cycles.


Background: The thioredoxin (trx) fusion E. Coli expression system is available to offer soluble expression of normally insoluble or difficult to express proteins. LaVallie et al. reported that a number of mammalian cytiokines and growth factors, when expressed as C-terminal trxA fusion proteins, stayed remarkably soluble in the E. Coli cytoplasm under certain conditions.


Mouse (Monoclonal) anti-Human Thioredoxin

cat# RDI-THIORDHabm    $500.00/100ug

Packaging: Protein A purified antibody from ascites, lyophilized. Reconstitute with 100ul distilled water to yield 1.0mg/ml stock IgG in PBS pH 7.2 with 1% sucrose, no preservative.

Isotype: mouse monoclonal IgG2a

Clone: 2E3

Immunogen: full length human thioredoxin fusion protein

APPLICATIONS : Western blotting; 1ug/ml (band approx 15kda) human:postive control=Jurkat, HeLa, Raji, U937 negative with mouse NIH3T3, WR19L, BaF/3

-not tested with other species

REFERENCE:

1) Holmgen, A. et al., Annu. Rev. Biochem. 54, 237-271 (1985)

2) Wollman, E. E. et al., J. Biol. Chem. 263, 15506-15512 (1988)

3) Sasada, T. et al., J. Toxicol. Sci. 21, 285-287 (1996)

4) Tanaka, H. et al., Vitam. Horm. 57, 153-175 (1999)

5) Powis, G. et al., Chem. Biol. Interact. 111-112, 2334 (1998)

BACKGROUND : Thioredoxin (TRX) is a small ubiquitous protein (MW12kDa) which is   exist in a wide variety of prokaryotic and eukaryotic cells. TRX   is activated by reducing its disulfide group (-S-S- .2SH)   between cysteine residues within the conserved active site   sequence : -Cys-Gly-Pro-Cys- . The reduction of TRX is specifically catalyzed by "TRX reductase", a FAD and selenocystein   containing protein, using NADPH. Once reduced, TRX in turn reduces disulfide bonds of various proteins to regulate the activity of the protein (redox regulation). TRX has multiple biological functions ; Activation of transcription factors such as NF-kB and glucocorticoid receptor, anti-apoptotic function, stimulation of cytokine expression, tissue protection from ischemia-reperfusion injury, cytoprotection from cytotoxic and DNA-damaging agents, proliferation, and drug resistance in certain malignancies. A part of these functions are due to TRX which exported out of the cell where it has additional ability to stimulate cell growth and cytoprotection, and co-cytokine activity. It is said that some of above TRX functions are related to response against the cellular "stresses". In fact, TRX istranslocated from the cytosol into the nucleus and/or highly expressed by various stresses such as UV irradiation and viral infection. So, it is suggested that TRX may have a role in the cellular responses against the stresses through the regulation of intracellular redox (oxidation-reduction) status

For Research Use Only

#RDI-THIORDH-AGX  $938.00/250ug recombinant human thioredoxin, from E. coli, free of E. coli S2, immunoaffinity purified, homogenous by SDS-PAGE. 250 µg vial, lyophilized.

#RDI-THIORDHabGX   $1062.00 goat anti-human thioredoxin IgG, immunoaffinity purified, 100 µg vial, lyophilized.

For In Vitro Research Use Only


Mouse anti-human THIOREDOXIN (full length, TRX 1-104)   MONOCLONAL ANTIBODY

cat# RDI-THIORDabm-11 $438.00

Presentation: 100ug in 0.2ml (0.5mg/ml) in 10 mM sodium phosphate, pH 7.2, with 150 mM NaCl and 0.09% (w/v) sodium azide.

Clone: 2G11/TRX

Isotype: Mouse IgG1
Immunogen: recombinant human Trx.

PREPARATION: purified from tissue culture supernatant via Protein G affinity

Reactivity: 2G11 reacts exclusively with Trx(1-104), but not with its truncated form.

Use: 1. Western Blotting: The 2G11 antibody is suitable to detect Trx (1-104) in western blots. Sensitivity: = 10 ng recombinant hu Trx, when BCIP/NBT substrate is used for blot development. 2. Immunofluorescent Staining and Flow Cytometric Analysis: A useful control for demonstrating specificity of staining with 2G11-FITC (Cat#RDI- THIORD-11FT $500.00/100ug). Pre-block the fixed/permeabilized cells with unlabeled 2G11 antibody prior to staining. The staining technique and blocking controls are described in detail by C. Prussin and D. Metcalfe. 4 A suitable mouse IgG 1 isotype control for assessing the level of background staining on paraformaldehyde- fixed/saponin-permeabilized human cells is FITC- MOPC-21 use at comparable concentrations to antibody of interest (e.g., =0.5 µg mAb/1 million cells).

DESCRIPTION

Thioredoxin (Trx) is an intracellular, multifunctional 12 kD (104a.a.) protein with reduction/oxidation (redox) activity expressed bymost cells of the human body. Trx is also secreted via the leaderless pathway (i.e. like IL-1 ) by cells such as monocytes and lymphocytes upon activation or in response to oxidative stress. A truncated, 10 kD form of Trx (Trx1-84) has also been reported. 1 Secreted Trx (also known as adult T- cell leukemia derived factor (ADF)) exhibits several cytokine- and chemokine-like functions such as activating eosinophils and granulocytes and chemotaxis for neutrophils, monocytes and T cells. 2,3

APPLICATION NOTES

Detection of Trx (1-104) by Western Blotting: recombinant huTrx (100, 50 25 or 12.5 ngs/lanes 1 - 4, respectively) or dilutions of THP-1 cell lysate (5x10 4 or 2.5x10 4 cell equivalent- - lanes 5 and 6 respectively) were separated on 14% SDS-PAGE under reducing conditions. The separeted proteins were transferred to PVDF membrane, and probed with 2G11 (1 µg/ml). AKP-conjugated polyclonal anti-mouse IgG was used as a secondary antibody at 1:1000 dilution, and the blot was developed with BCIP/NBT substrate (Sigma).

The 2G11 antibody detects a single, approx. 12 kDa band protein in lanes 1 - 4 (rhuTrx) as well as well as in the cellular lysate, that corresponds to the MW of Trx. . Purified mouse IgG1k is recommended as an isotype control

REFERENCES

1. Sahaf, B., A. Soderberg, G. Spyrou, A. M. Barral, K. Pekkari, A. Holmgren and A. Rosen. 1997. Thioredoxin expression and localization in human cell lines: Detection of full length and truncated species. Exp. Cell Research . 236: 181-192.

2 Bertini, R., O. M. Z. Howard, H-F.Dong, J. J. Oppenheim, C. Bizzarri, R. Sergi, G. Caselli, S. Pagliei, B. Romines, J.A. Wilshire, M.Mengozzi, H. Nakamura, J. Yodoi, K. Pekkari, R. Gurunath, A. Holmgren, L. A. Herzenberg, L. A. Herzenberg, and P. Ghezzi. 1999. Thioredoxin, a redox enzyme released in infection and inflammation, is a unique chemoattractant for neutrophils monocytes and T cells. J. Exp. Med. 189:1783-1789.

3. Yoshida, S., T. Katoh, T. Tetsuka, K. Uno, N. Matsui and T. Okamoto. 1999. Involvement of thioredoxin in rheumatoid arthritis: Its costimulatory roles in the TNF- - induced production of IL-6 and IL-8 from cultured synovial fibroblast. J .Immunol. 163:351-358.

4. Prussin, C., and D. Metcalfe. 1995. Detection of intracytoplasmic cytokine using flow cytometry and directly conjugated anti-cytokine antibodies. J. Immunol. Meth. 188:117- 28.

5 Immunofluorescent staining of intracellular cytokines for flow cytometric analysis. 1998. In Cytokine/Chemokine Manual: Genes to Proteins to Cells. Application Manual. 3rd Edition.

For Research Use Only. Not For Diagnostic or Therapeutic Use.

Conditions: Not responsible for patent infringement or other violations that may occur with the use of our products.

Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.


RDI Divison of Fitzgerald Industries Intl

34 Junction Square Drive

Concord MA 01742-3049

USA

phone (800) 370-2222

     or  (978) 371-6446 or (800) 370-2222

fax     (978) 371-2266

EMAIL:antibodies@fitzgerald-fii.com

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